An enzymatic method for the measurement of glycated albumin in biological samples

Clin Chim Acta. 2002 Oct;324(1-2):61-71. doi: 10.1016/s0009-8981(02)00207-3.


Background: In order to determine glycated albumin more easily and rapidly, we developed a new enzymatic method for glycated albumin in blood samples.

Methods: The method involves use of albumin-specific proteinase, ketoamine oxidase and serum albumin assay reagent. In the assay, glycated albumin is hydrolyzed to glycated amino acids by proteinase digestion, and ketoamine oxidase oxidizes the glycated amino acids to produce hydrogen peroxide, which is quantitatively measured. Glycated albumin is calculated as the percentage of glycated albumin in total albumin.

Results: The calibration curve for glycated albumin concentration was linear (r(p)=0.999) between 0.0 and 50.0 g/l and that for albumin concentration was linear (r(p)=0.999) between 0.0 and 60.0 g/l. The analytical recoveries of exogenous glycated albumin added to serum were 100-102.5%. The within-run and between-run CVs were 0.45-0.67% and 1.09-1.26%, respectively. This method was free from interference by bilirubin, chyle, glucose, globulins and labile intermediate. Weak interference by hemoglobin and ascorbic acid was observed. Glycated albumin detected by the present method was significantly correlated with glycated albumin detected by high-performance liquid-chromatographic (HPLC) method (serum: r(s)=0.989, plasma: r(p)=0.992).

Conclusions: This new enzymatic method is simple, rapid, allows multiple determinations and enables quantitative analysis of glycated albumin.

MeSH terms

  • Amino Acids / analysis
  • Amino Acids / chemistry
  • Calibration
  • Chromatography, High Pressure Liquid
  • Detergents
  • Diabetes Mellitus / blood
  • Endopeptidases / metabolism*
  • Humans
  • Hyperglycemia / blood
  • Oxidoreductases / metabolism
  • Protein Binding
  • Sensitivity and Specificity
  • Serum Albumin / analysis*
  • Substrate Specificity
  • Time Factors


  • Amino Acids
  • Detergents
  • Serum Albumin
  • glycosylated serum albumin
  • Oxidoreductases
  • Endopeptidases